Application of N-acetylneuraminic acid substances to regulation of intestinal flora of mother and child

Abstract

The invention belongs to the field of food, and particularly relates to application of N-acetylneuraminic acid substances to regulation of intestinal flora of a mother and a child. The invention provides an application of N-acetylneuraminic acid substances in preparation of food which is taken by a mother and contributes to regulation of intestinal flora of the mother and filial generations of the mother; wherein the N-acetylneuraminic acid substance is an N-acetylneuraminic acid anhydride, an N-acetylneuraminic acid hydrate or an N-acetylneuraminic acid salt, the mother is in a gestation period and / or a lactation period, and the offspring is in a fetal period, an infant period and / or a children period. The invention accidentally finds that when a mother starts to ingest N-acetylneuraminic acid substances in the gestation period, the increase of beneficial bacteria in intestinal tracts in the gestation period and the lactation period can be promoted, meanwhile, the content of probiotics in offspring intestinal tracts can be increased, the proliferation of conditioned pathogenic bacteria in the offspring intestinal tracts is inhibited, and the relative abundance of the conditioned pathogenic bacteria is reduced; and the health of the nervous system is further maintained through the effect of the intestinal brain axis.

Description

Technical Field

[0001] The invention belongs to the field of food, and specifically relates to the use of N-acetylneuraminic acid substances in helping to regulate the intestinal flora of mother and child. Background Art

[0002] There are a large number of complex microorganisms in the intestine, an important digestive system of humans, which have co-evolved with the host. Existing studies have found that the main members of the human intestinal flora are Bacteroidetes, Firmicutes, Actinobacteria, Proteobacteria, Verrucomicrobia, etc. The intestinal flora is called the "second brain" of humans, and a large number of studies have found that the intestinal flora plays an important role in human health. The first 1000 days of life is not only a critical period for brain development, but also a critical period for the establishment of intestinal flora. The establishment of a healthy intestinal flora in early life has an important and far-reaching impact on the health outcomes of humans from the fetal period, childhood to adulthood.

[0003] Studies have shown that the nutritional supply of the mother to the fetus during pregnancy has an important impact on the health outcomes of the offspring throughout their life cycle. Furthermore, during the lactation stage, breast milk can not only provide nutrients to the fetus, but also provide the fetus with immune active components and beneficial microorganisms that are important for its healthy growth and development. Existing studies believe that the initial flora in the gastrointestinal tract of newborns comes from the mother. The mother's healthy flora can pass on better flora to the offspring and affect the establishment of the offspring's flora, laying a good foundation for the healthy establishment of the offspring's flora.

[0004] N-acetylneuraminic acid is a nine-carbon monosaccharide with an N-acetylamino group, also known as neuraminic acid, and is a common sialic acid. N-acetylneuraminic acid is usually fused to the non-reducing ends of glycolipids and glycoproteins on the cell membrane, and is involved in cell recognition and the regulation of various biological processes. Studies have found that sialic acid has the function of regulating the anti-inflammatory activity of IgG, can enhance the body's immunity, and affect the integrity, permeability and activity of nerve cells. At present, N-acetylneuraminic acid can be used as a raw material in food.

[0005] Reference document 1 (CN102946728B) discloses a method for providing sialic acid to a woman, comprising administering sialic acid in the form of N-acetylneuraminic acid to the woman during at least one of the stages of pre-pregnancy, pregnancy and lactation, wherein the sialic acid provides developmental benefits to the nervous system and / or brain of the fetus or child of the woman.

[0006] Reference 2 (CN119054770A) discloses that sialic acid, as a nutrient for intestinal microorganisms, can promote the growth of microorganisms with sialic acid metabolism ability, such as Bacteroides fragilis and Akkermansia, and affect intestinal homeostasis. Studies have shown that supplementing sialic acid-containing glycoproteins or sialylated glycans can change the composition of the entire intestinal microbiota, thereby improving the health of the body, including reducing the risk of colorectal cancer and atherosclerosis. Intake of sialic acid from the diet, especially conjugated sialic acid glycans, is particularly important for human intestinal health. Summary of the invention

[0007] Problem that the invention aims to solve

[0008] The mother's overall nutritional environment during pregnancy and lactation affects the nutrients she provides to her offspring, but there are currently few studies on improving maternal nutrition to regulate the establishment of her offspring's intestinal flora.

[0009] Existing studies have shown that supplementing N-acetylneuraminic acid during pregnancy can benefit the development of the fetal nervous system, but no study has ever shown that supplementing N-acetylneuraminic acid during pregnancy can regulate the maternal intestinal flora and further regulate the intestinal flora of the offspring. Although, for example, the above-mentioned reference 2 discloses the effect of sialic acid on intestinal flora, on the one hand, its research focuses on sialic acid glycans, and on the other hand, even if nutrients are believed to be directly given to the body to exert specific health benefits, they may not necessarily promote the health of the offspring when consumed by the mother.

[0010] In the process of conducting in-depth research on the physiological functions of N-acetylneuraminic acid substances, the present invention unexpectedly discovered that mothers who begin to ingest N-acetylneuraminic acid substances during pregnancy can significantly improve their intestinal flora and can also improve the intestinal flora of their offspring at the same time. N-acetylneuraminic acid substances can promote the increase of beneficial bacteria in the intestines of pregnant and lactating mothers, and can also increase the content of probiotics in the intestines of their offspring, inhibit the proliferation of conditional pathogens in the intestines of the offspring and reduce their relative abundance.

[0011] Therefore, the object of the present invention is to provide a non-therapeutic use of N-acetylneuraminic acid substances in simultaneously regulating the intestinal flora of pregnant and / or lactating mothers and their offspring. Further, the object of the present invention is to provide a use of N-acetylneuraminic acid substances in preparing foods that help regulate the intestinal flora of mothers and their offspring through ingestion by mothers.

[0012] Solutions for solving problems

[0013] [1]. Use of N-acetylneuraminic acid substances in the preparation of foods that help regulate the intestinal flora of the mother and her offspring through ingestion; wherein the N-acetylneuraminic acid substance is N-acetylneuraminic acid anhydrate, N-acetylneuraminic acid hydrate or N-acetylneuraminic acid salt, the mother is in pregnancy and / or lactation, and the offspring is in the fetal period, infancy, and / or childhood.

[0014] [2] The method according to [1], wherein the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Muribaculaceae unclassified in the intestinal tract of pregnant mothers, increasing the relative abundance of Prevotellaceae UCG-001 in the intestinal tract of pregnant mothers, and increasing the relative abundance of Bifidobacterium in the intestinal tract of pregnant mothers.

[0015] [3] The use according to [1] or [2], wherein the method for helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of Muribaculaceae unclassified in the intestine of the mother during lactation and increasing the relative abundance of Akkermansia in the intestine of the mother during lactation.

[0016] [4] The use according to any one of [1] to [3], wherein the method for helping to regulate the intestinal flora of the mother and her offspring comprises: increasing the relative abundance of Muribaculaceae unclassified in the offspring's intestine, increasing the relative abundance of Akkermansia in the offspring's intestine, increasing the relative abundance of Romboutisa in the offspring's intestine, and decreasing at least one of the relative abundance of Escherichia-Shigella in the offspring's intestine.

[0017] [5]. The use according to any one of [1] to [4], wherein the food affects the gut-brain axis of the mother and her offspring by helping to regulate their intestinal flora, thereby helping to maintain the health of the nervous system of the mother and her offspring.

[0018] [6]. The use according to any one of [1] to [5], wherein the food is infant food, children's food, adolescent food or adult food.

[0019] [7]. The use according to any one of [1] to [6], wherein the food is a beverage, candy, dairy product, baked food or dietary supplement.

[0020] [8]. The use according to any one of [1] to [7], wherein the food is in liquid form, solid-liquid mixed form, solid block form or solid powder form at room temperature.

[0021] [9]. The use according to any one of [1] to [8], wherein the food contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional additives and any excipients acceptable to food.

[0022]

[10] . The use according to any one of [1] to [9], wherein the mass content of the N-acetylneuraminic acid in the food is 0.03% to 2.0%.

[0023] Effects of the Invention

[0024] After extensive research, the present invention proposes that supplementing a certain amount of N-acetylneuraminic acid substances during pregnancy and / or lactation can significantly increase the abundance of beneficial bacteria in the intestines of mothers, and at the same time significantly increase the abundance of beneficial bacteria in the intestines of their offspring and reduce the content of harmful bacteria, especially regulating the abundance of some non-directly edible beneficial bacteria in the intestines of mothers and offspring. Furthermore, regulating the intestinal flora will affect the gut-brain axis of the body. Therefore, the present invention proposes that supplementing a certain amount of N-acetylneuraminic acid substances during pregnancy and / or lactation is also beneficial to maintaining the health of the nervous system of the mother and her offspring.

[0025] Experimental data show that supplementary intake of N-acetylneuraminic acid substances by mothers during pregnancy and lactation can significantly increase the relative abundance of beneficial bacteria Muribaculaceae unclassified, Prevotellaceae UCG-001 and Bifidobacterium in the mother's intestines during pregnancy; increase the relative abundance of beneficial bacteria Muribaculaceae unclassified and Akkermansia in the mother's intestines during lactation; at the same time, it can significantly increase the relative abundance of beneficial bacteria Muribaculaceae unclassified, Akkermansia and Romboutisa in the intestines of the mother's offspring, and reduce the relative abundance of conditionally pathogenic bacteria Escherichia-Shigella in the intestines of the mother's offspring. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 : Changes of Muribaculaceae unclassified in the intestine of female mice.

[0027] Figure 2 :Changes of Prevotellaceae UCG-001 in the intestines of female mice.

[0028] Figure 3 :Changes of Bifidobacterium in the intestine of female mice.

[0029] Figure 4 :Changes of Akkermansia in the intestines of female mice.

[0030] Figure 5 : Changes of Muribaculaceae unclassified in the intestine of offspring mice.

[0031] Figure 6 :Changes of Akkermansia in the intestine of offspring mice.

[0032] Figure 7 :Changes of Romboutisa in the intestine of offspring mice.

[0033] Figure 8 :Changes of Escherichia-Shigella in the intestine of offspring mice. DETAILED DESCRIPTION

[0034] The following describes the embodiments of the present invention, but the present invention is not limited thereto. Various changes can be made within the scope of the present invention, and the embodiments and examples obtained by appropriately combining the technical means disclosed in different embodiments and examples are also included in the technical scope of the present invention.

[0035] I. Definition of Terms

[0036] In the present invention, the term "a" or "an" or "the" may mean "one", and may also mean "one or more", "at least one" and "one or more than one".

[0037] In the present invention, the numerical ranges expressed using “value A to value B”, “value A - value B”, “above value A”, “below value A”, “at least value A” and “at most value A” refer to the range including the endpoint values ​​A and B.

[0038] In the present invention, "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes instances where the event occurs and instances where it does not occur.

[0039] In the present invention, the word "may" includes both performing a certain process and not performing a certain process.

[0040] In the present invention, the terms "comprising", "having", "including" or "containing" may mean inclusive or open-ended, and do not exclude additional, uncited elements or method steps. At the same time, "comprising", "having", "including" or "containing" may also mean closed, excluding additional, uncited elements or method steps.

[0041] In the present invention, the term "about" is used to define the numerical range and parameters of the present invention are approximate values, and the specific relevant values ​​are presented as accurately as possible. Unless otherwise expressly stated, it should be understood that all ranges, quantities, values ​​and percentages used in the present invention are modified by "about". Here, "about" usually means that the actual value is within ±5%, ±3%, ±1% or ±0.5% of a particular value or range.

[0042] In the present invention, "normal temperature" refers to an indoor ambient temperature of 23±2°C.

[0043] In the present invention, "animal milk" is used to refer to the liquid obtained from the mammary gland of a mammal in the lactation period. The term "animal milk" should be interpreted broadly and covers both raw milk (ie, liquid obtained directly from the mammary gland) and standardized milk products.

[0044] In the present invention, "gestation period" and "pregnancy period" are used interchangeably and refer to the period from fertilization to birth.

[0045] In the present invention, the "breastfeeding period" refers to the period from the start of breastfeeding to the cessation of breastfeeding after a mother gives birth.

[0046] In the present invention, "infants" refers to a human group under 36 months of age.

[0047] In the present invention, "infant" refers to a human group under 12 months of age.

[0048] In the present invention, "infants" refers to a human group of 13 to 36 months old.

[0049] In the present invention, "children" refers to a human group older than 3 years and younger than 12 years.

[0050] In the present invention, "teenagers" refers to a human group aged 12 years or older and younger than 18 years old.

[0051] In the present invention, "adult" refers to a human group of people aged 18 years or older.

[0052] In the present invention, "middle-aged and elderly" refers to a human group over the age of 45.

[0053] Unless otherwise defined, other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0054] II. N-acetylneuraminic acid

[0055] N-acetylneuraminic acid (Neu5Ac) is a common form of sialic acid and an acetylated derivative of acidic sugar neuraminic acid. It plays an important role in biological recognition, cellular immunity and disease, and is one of the most important monomers of sialylated human milk oligosaccharides.

[0056] The present invention does not particularly limit the source of N-acetylneuraminic acid substances. Typically, they can be extracted from natural resources containing sialic acid, such as bird's nests, eggs, milk, etc.; or, they can be synthesized by chemical and enzymatic methods, such as condensing N-acetylglucosamine with potassium salt of di-tert-butyloxysuccinic acid, and decarboxylating under the catalysis of alkali to generate N-acetylneuraminic acid, or using N-acetylneuraminic acid aldolase to catalyze N-acetylmannosamine for production; or, they can be obtained by fermentation with suitable microorganisms such as Escherichia coli and Bacillus subtilis; or, they can be obtained by whole-cell synthesis.

[0057] In some embodiments, the N-acetylneuraminic acid substance of the present invention is N-acetylneuraminic acid anhydride (abbreviated as N-acetylneuraminic acid), N-acetylneuraminic acid hydrate or N-acetylneuraminic acid salt; preferably N-acetylneuraminic acid anhydride or N-acetylneuraminic acid hydrate; more preferably N-acetylneuraminic acid anhydride. Exemplary N-acetylneuraminic acid hydrates include N-acetylneuraminic acid dihydrate, which has basically the same biological efficacy as N-acetylneuraminic acid. The two have the same properties except that there is a little difference in the solid state due to the presence of crystal water. In the liquid state, the two have the same properties due to the break of the hydrogen bonds connecting the two crystal waters of the dihydrate to N-acetylneuraminic acid. Exemplary N-acetylneuraminic acid salts include N-acetylneuraminic acid sodium salt, N-acetylneuraminic acid potassium salt or N-acetylneuraminic acid ammonium salt. For the preparation method of N-acetylneuraminic acid salt, the present invention is not limited, for example, a solvent composed of ketones or alcohols can be added to an N-acetylneuraminic acid aqueous solution containing sodium, potassium or ammonium at a pH of 3.0 to 9.0, and finally crystals are precipitated.

[0058] In some specific embodiments, the N-acetylneuraminic acid substances of the present invention can be prepared by themselves or purchased commercially.

[0059] II. Foods containing N-acetylneuraminic acid (including health foods)

[0060] The food containing N-acetylneuraminic acid substances of the present invention is suitable for people of all ages, especially for people who need to regulate intestinal flora, and is particularly suitable for mothers in pregnancy and / or lactation. In some specific embodiments, the food of the present invention is infant food, children's food, adolescent food or adult food; exemplary adult food includes pregnant women food, maternal food, maternal food, middle-aged and elderly food, etc.

[0061] The food containing N-acetylneuraminic acid substances described in the present invention can be any kind of food. In some specific embodiments, the food described in the present invention is a beverage, candy, dairy product, baked food or dietary supplement; exemplary beverages include carbonated beverages, tea beverages, coffee beverages, fruit and vegetable juice beverages and lactic acid bacteria beverages, etc.; exemplary candies include hard candies, gel candies, crisp candies, pressed candies and inflated candies, etc.; exemplary dairy products include fermented milk, cheese and milk powder, etc.; exemplary baked food includes bread, cakes and biscuits, etc.; exemplary dietary supplements include hard capsules, soft capsules, tablets, oral liquids, granules and powders, etc.

[0062] The food containing N-acetylneuraminic acid substances of the present invention can be in any form. In some specific embodiments, at room temperature, the food of the present invention is in liquid form, solid-liquid mixed form, solid block form or solid powder form.

[0063] In the food of the present invention, in addition to containing N-acetylneuraminic acid substances, other arbitrary food acceptable substances may also be included, such as protein / amino acid, carbohydrate, fat, vitamin, mineral, etc., and those skilled in the art can select according to the actual food type and target population. In some specific embodiments, the food of the present invention contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional added ingredients and any acceptable auxiliary materials in food. Exemplary plant product ingredients include various vegetables, fruits, grains, nuts, coffee, tea and their extracts, as well as some medicinal and edible botanical Chinese medicinal materials and their extracts. Exemplary animal dairy product ingredients include fresh milk from animals, as well as whole milk powder, skim milk powder, concentrated whey protein powder, desalted whey powder, whey protein powder, hydrolyzed whey protein powder, casein powder and other reprocessed dairy products. Exemplary animal meat product ingredients include meat product ingredients of pigs, cattle, sheep, aquatic products or poultry. Exemplary functional added ingredients include vitamin supplements, mineral supplements, nucleotide supplements, dietary fiber, functional polyunsaturated fatty acid supplements, etc. Exemplary acceptable excipients in foods include solvents, antioxidants, antimicrobial agents, thickeners, diluents, cosolvents, stabilizers, emulsifiers, fillers, disintegrants, lubricants, coating materials, anti-caking agents, flavoring agents, sweeteners, edible flavors, edible coloring, etc.

[0064] In some more specific embodiments, the food of the present invention is infant formula milk powder, baby complementary food, children's formula milk powder, children's snacks, maternal formula milk powder, pregnant woman formula milk powder, middle-aged and elderly milk powder, maternal oral liquid or pregnant woman oral liquid.

[0065] In the food of the present invention, the content of N-acetylneuraminic acid substances meets the requirements of relevant laws and regulations. In some specific embodiments, in the food of the present invention, the mass content of the N-acetylneuraminic acid substances is 0.03% to 2.0%, for example, it can be 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9% or 2.0%, etc., preferably 0.08% to 1.3%.

[0066] III. Use of the drug to help regulate the intestinal flora of the mother and her offspring through maternal ingestion

[0067] The present invention finds that a mother's intake of a certain amount of N-acetylneuraminic acid substances during pregnancy and / or lactation (consumption) can help regulate the intestinal flora of the mother and her offspring, which is specifically manifested in that the relative abundance of beneficial bacteria in the intestinal tract of the mother and her offspring can be increased, and the relative abundance of conditional pathogens in the intestinal tract of the offspring can be reduced. At the same time, the present invention finds that the effect of N-acetylneuraminic acid substances ingested by the mother to regulate the intestinal flora of the offspring is not only for the fetus present in the mother's body and infants who consume breast milk, but also for infants who do not consume breast milk but are in full contact with the mother. At the same time, this regulatory effect can also continue to the childhood of the offspring, and even have a lasting effect on the offspring.

[0068] Based on this, the present invention provides the non-therapeutic use of N-acetylneuraminic acid substances and foods containing N-acetylneuraminic acid substances, which help regulate the intestinal flora of the mother and her offspring through ingestion by the mother. At the same time, the present invention provides the use of N-acetylneuraminic acid substances in the preparation of foods that help regulate the intestinal flora of the mother and her offspring through ingestion by the mother.

[0069] In some embodiments, the mother is in pregnancy and / or lactation; preferably, the mother is in pregnancy and lactation. In some embodiments, the offspring is in the fetal period, infant period, and / or childhood; preferably, the offspring is in the fetal period and / or infant period and optional childhood; more preferably, the offspring is in the infant period and optional childhood; even more preferably, the offspring is in the infant period. In some embodiments, the offspring in the infant period can be fed with breast milk, preferably completely fed with breast milk or mainly fed with breast milk. In the present invention, "completely fed with breast milk" means that the vast majority (at least 90%, preferably at least 95%, more preferably at least 98%) of the nutrients and / or energy consumed by the offspring comes from breast milk. In the present invention, the use of "mainly fed with breast milk" means that the nutrients and / or energy consumed by the offspring mainly (at least 50%, preferably at least 65%, more preferably at least 75%) come from breast milk.

[0070] In some embodiments, the "mother" and "offspring" described in the present invention include mammals, including but not limited to humans, monkeys, gorillas, cows, sheep, cats, dogs, horses, rabbits, mice, rats or guinea pigs, etc.

[0071] The present invention finds that although N-acetylneuraminic acid substances are ingested by mothers, the intestinal regulatory effects of N-acetylneuraminic acid substances on mothers and offspring at different stages are not exactly the same. In some embodiments, supplementary intake of N-acetylneuraminic acid substances by mothers during pregnancy and / or lactation can regulate different flora in the intestines of pregnant mothers, lactating mothers and their offspring, respectively.

[0072] In some embodiments, the method of helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of beneficial bacteria in the intestine of the pregnant mother, increasing the relative abundance of beneficial bacteria in the intestine of the lactating mother, increasing the relative abundance of beneficial bacteria in the intestine of the offspring, and reducing the relative abundance of conditional pathogens in the intestine of the offspring.

[0073] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of beneficial bacteria in the intestine of pregnant mothers, increasing the relative abundance of beneficial bacteria in the intestine of lactating mothers, increasing the relative abundance of beneficial bacteria in the intestine of offspring, and reducing the relative abundance of conditional pathogens in the intestine of offspring.

[0074] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Muribaculaceae unclassified in the intestinal tract of the pregnant mother, increasing the relative abundance of Prevotellaceae UCG-001 in the intestinal tract of the pregnant mother, and increasing the relative abundance of Bifidobacterium in the intestinal tract of the pregnant mother.

[0075] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Muribaculaceae unclassified in the intestinal tract of the pregnant mother, increasing the relative abundance of Prevotellaceae UCG-001 in the intestinal tract of the pregnant mother, and increasing the relative abundance of Bifidobacterium in the intestinal tract of the pregnant mother.

[0076] In some embodiments, the method of helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of Muribaculaceae unclassified in the intestinal tract of the lactating mother and increasing the relative abundance of Akkermansia in the intestinal tract of the lactating mother.

[0077] In some embodiments, the helping to regulate the intestinal flora of the mother and her offspring comprises: increasing the relative abundance of Muribaculaceae unclassified in the intestinal tract of the lactating mother and increasing the relative abundance of Akkermansia in the intestinal tract of the lactating mother.

[0078] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Muribaculaceae unclassified in the offspring's intestine, increasing the relative abundance of Akkermansia in the offspring's intestine, increasing the relative abundance of Romboutisa in the offspring's intestine, and reducing at least one of the relative abundance of Escherichia-Shigella in the offspring's intestine.

[0079] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Muribaculaceae unclassified in the offspring's intestine, increasing the relative abundance of Akkermansia in the offspring's intestine, increasing the relative abundance of Romboutisa in the offspring's intestine, and reducing the relative abundance of Escherichia-Shigella in the offspring's intestine.

[0080] Muribaculaceae unclassified is an unnamed genus under the family Muribaculaceae. Muribaculaceae is a bacterial family in the order Bacteroidetes. Muribaculaceae produces short-chain fatty acids through endogenous (mucin polysaccharides) and exogenous polysaccharides (dietary fiber). In recent years, Muribaculaceae has begun to be paid attention to and studied as a potential probiotic family. Studies have found that this family has a cross-feeding relationship with probiotics such as Bifidobacterium and Lactobacillus. Muribaculaceae can effectively decompose dietary fiber into monosaccharides and then serve as a food source for Bifidobacterium and Lactobacillus to promote the proliferation of Lactobacillus and Bifidobacterium (Zhu Y, Chen B, Zhang X, Akbar MT, Wu T, Zhang Y, ZhiL, Shen Q. Exploration of the Muribaculaceae Family in the Gut Microbiota: Diversity, Metabolism, and Function. Nutrients. 2024Aug12; 16(16): 2660.).

[0081] Prevotellaceae UCG-001 is generally considered to be a bacterium associated with a healthy plant-based diet. Studies have shown that Prevotellaceae UCG-001 is positively correlated with the AMPK signaling pathway in the body and negatively correlated with abnormal glucose and lipid metabolism. Studies have also shown that the level of Prevotellaceae in the intestines of patients with obsessive-compulsive disorder and depression is reduced (Borrego-Ruiz A, Borrego JJ. An updated overview on the relationship between human gut microbiome dysbiosis and psychiatric and psychological disorders. Prog Neuropsychopharmacol Biol Psychiatry. 2024Jan10; 128: 110861.).

[0082] Bifidobacterium microorganisms are Gram-negative anaerobic bacteria in the intestine. Studies have shown that Bifidobacterium microorganisms have a variety of probiotic functions, such as improving intestinal diseases caused by immune system disorders; inhibiting pathogens from invading the intestine; improving the activity of host antioxidant enzymes to alleviate oxidative damage to the body; improving diseases caused by intestinal flora imbalance, such as constipation and diarrhea; secreting β-galactosidase to alleviate lactose intolerance, etc. Studies have found that Bifidobacterium microorganisms have effects on memory and learning behavior, long-term potentiation (LTP) and some biochemical parameters in β-Amyloid-Induced Rat's Model of Alzheimer's Disease. Prev Nutr Food Sci. 2019 Sep; 24 (3): 265-273.).

[0083] Akkermansia microorganisms are Gram-negative anaerobic bacteria in the intestine. They mainly colonize in the mucus layer in the gastrointestinal tract and protect the mucus layer and connective tissue (Derrien M, Vaughan EE, Plugge CM, de Vos WM. Akkermansia muciniphila gen. nov., sp. nov., a human intestinal mucin-degrading bacterium. Int J Syst Evol Microbiol. 2004 Sep; 54 (Pt5): 1469-1476.); as a new generation of probiotics, they are a biomarker of health status (Zhang T, Li Q, Cheng L, Buch H, Zhang F. Akkermansia muciniphila is a promising probiotic. Microb Biotechnol. 2019 Nov; 12 (6): 1109-1125.). Studies have shown that Akkermansia microorganisms play an important role as probiotics in metabolic diseases (such as diabetes, obesity, etc.), intestinal diseases (such as inflammatory bowel disease, ulcerative colitis, etc.) and neuropsychiatric systems (Huang Qiaoshen, Zhang Yongkang, Wang Shuiping, et al. Research progress on the potential and mechanism of action of Akkermansia muciniphila in treating diseases [J]. Journal of Microbiology, 2023, 63(09): 3360-3373.). With the continuous deepening of research on the gut-brain axis, studies have found that Akkermansia plays an important role in a variety of neuropsychiatric diseases (such as depression and anxiety, autism spectrum disorders (ASDs), Alzheimer's disease and cognitive impairment, etc.).

[0084] Romboutisa is an important cornerstone bacteria in the human intestine and is part of the symbiotic bacteria. It can use a variety of carbohydrates to produce short-chain fatty acids such as formic acid, acetic acid, lactic acid and butyric acid, ferment single amino acids, ferment to produce vitamins, affect colon motility, and has anti-inflammatory properties. Studies have shown that the relative abundance of Romboutisa in the intestine of rats in a depressed state is significantly reduced (Wang Qiannan, Huang Xinhui, Yang Minxu, et al. Research progress on antidepressant treatment related to intestinal flora [J]. Pharmaceutical Practice and Service, 2022, 40(05): 422-426.).

[0085] Escherichia-Shigella is a genus of the Enterobacteriaceae family. It is one of the common pathogenic bacteria in the intestine. It is a Gram-negative facultative anaerobic bacterium. Its growth does not require high nutrition. It can produce enterotoxins with high pathogenicity. Its large-scale reproduction can cause intestinal flora disorders and cause bacillary dysentery. Studies have shown that Escherichia-Shigella is more abundant in patients with generalized anxiety disorder and is positively correlated with the severity of anxiety (Chen YH, Bai J, Wu D, Yu SF, Qiang XL, Bai H, Wang HN, Peng ZW. Association between fecal microbiota and generalized anxiety disorder: Severity and early treatment response. J Affect Disord. 2019 Dec 1; 259: 56-66.).

[0086] Based on the experimental results of the present invention and existing literature reports, the present invention believes that by allowing the mother to supplement the intake of N-acetylneuraminic acid substances during pregnancy and / or lactation, it is not only beneficial to the health of the intestinal flora of the mother and her offspring, but also may affect the gut-brain axis of the mother and her offspring, thereby having a certain beneficial effect on the nervous system health of the mother and her offspring. The invention's help in maintaining the nervous system health of the mother and her offspring is not intended to prevent and treat diseases.

[0087] The present invention has found through a large number of studies that supplementary intake of N-acetylneuraminic acid substances by mothers during pregnancy and / or lactation can simultaneously regulate some of the same flora in the intestines of pregnant and / or lactating mothers and their offspring, especially some probiotics that are not on the list of regional edible probiotics and cannot be directly supplemented exogenously for the time being.

[0088] In some embodiments, the method helps to regulate the intestinal flora of the mother and her offspring, including: simultaneously increasing the relative abundance of Muribaculaceae unclassified in the intestinal tract of the mother and her offspring during pregnancy and lactation.

[0089] In some embodiments, the method of helping to regulate the intestinal flora of the mother and her offspring includes: simultaneously increasing the relative abundance of Akkermansia in the intestinal tract of the mother and her offspring during lactation.

[0090] Example

[0091] The embodiments of the present invention will be described in detail below in conjunction with examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out under conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the materials or instruments used are all conventional products that can be obtained commercially.

[0092] 1. Test materials

[0093] 1.1. Experimental animals and groups

[0094] 40 8-week-old sexually mature SPF female rats and 40 SPF male rats were selected; purchased from Hangzhou Medical College, animal certificate number 20240318Aazz0100000836, production license number: SCXK (Zhejiang) 2024-002. Feeding conditions: Animals were raised in a barrier environment, with an indoor temperature of 25±1℃, humidity of 45±5%, free access to drinking water, and 12h light and dark alternation every day. This experiment passed the animal experiment ethics review of Southeast University, ethics number: 20240316003.

[0095] After 3 days of animal adaptive feeding, female and male rats were caged in a 1:1 ratio. The intervention was started by gavage when the mother rat showed vaginal plugs, and the mother rats were continuously gavaged until the offspring rats were weaned. The intervention group was divided into groups and given the corresponding dose of sample aqueous solution, and the control group was given the corresponding dose of normal saline, gavaged once a day for 6 consecutive weeks. All groups were given maintenance feed, and the low and high doses of N-acetylneuraminic acid in the intervention group were 1.5 and 7.5 (mg / d / rat). The specific doses of each group are shown in Table 1 below. The feces of the mother rats before gavage, after delivery, and after weaning, and the feces of the offspring rats one week and three weeks after birth (after weaning) were collected for intestinal flora sequencing.

[0096] Table 1 Oral doses for experimental animals

[0097] Group Dosage Group N-acetylneuraminic acid mg / d / each Blank control Basic feed + saline Example 1 Basic feed + low dose of N-acetylneuraminic acid 1.5 Example 2 Basic feed + high dose of N-acetylneuraminic acid 7.5

[0098] 1.2. Instruments, consumables and reagents

[0099] The consumables, reagents and experimental instruments used in the experiment are shown in Tables 2 and 3.

[0100] Table 2 Main experimental consumables and reagents

[0101]

[0102] Table 3 Main experimental instruments

[0103] Instrument Name company Electronic balance PTX-FA110S Huazhi (Fujian) Electronic Technology Co., Ltd. Desktop high speed refrigerated centrifuge Xiangyi Agilent 2100 Bioanalyzer Agilent Qubit Fluorometer Invitrogen NovaSeq 6000 Illumina

[0104] 2. Detection Methods

[0105] 2.1. Feces collection of mother mice and offspring

[0106] (1) Feces collection of maternal mice: Fresh feces of each maternal mouse were collected in a clean environment before oral gavage, after delivery, and after weaning. The feces were numbered and stored at -80°C.

[0107] (2) Collection of feces of offspring: Fresh feces of each litter were collected at 1 week and 3 weeks after birth (after weaning), numbered, and stored in a frozen state at -80°C.

[0108] 2.2. Detection of fecal flora of mother mice and offspring

[0109] High-throughput sequencing of microorganisms in fecal samples of mother mice and offspring mice using 16S rDNA:

[0110] (1) Extraction of total DNA of the microbiome: A DNA extraction kit was used to extract the total DNA of the microbiome in the feces, and the quality of DNA extraction was tested by agarose gel electrophoresis. The DNA was quantified using an ultraviolet spectrophotometer.

[0111] (2) PCR amplification: The V3-V4 region of bacterial 16s rDNA was selected for gene amplification and sequencing. The primer sequences are shown in the following table:

[0112] Table 4 Primer sequences

[0113]

[0114] PCR reaction system is shown in Table 5 below:

[0115] Table 5 Reaction system

[0116] PCR reaction components PCR reaction volume Phusion Hot Start Flex 2X Master Mix 12.5μL Forward primer 2.5μL Reverse primer 2.5μL Template DNA 50ng <![CDATA[Add ddH2O to]]> 25μL

[0117] PCR reaction conditions Table 6:

[0118] Table 6 Reaction conditions

[0119]

[0120] (3) PCR products were purified using AMPure XT beads (Beckman Coulter Genomice, MA, USA) and quantified using Qubit (Invitrogen, USA).

[0121] (4) The purified PCR products were evaluated using the Agilent 2100 bioanalyzer (Agilent, USA) and the Illumina (Kapa Biosciences, Wobum, MA, USA) library quantification kit. The qualified library concentration was above 2 nM. The qualified libraries for sequencing were gradient diluted, mixed according to the required sequencing amount, and denatured into single strands by NaOH for sequencing. The NovaSeq 6000 sequencer was used for 2×250 bp double-end sequencing, and the corresponding reagent was the NovaSeq6000SP Reagent Kit (500 cycles).

[0122] (5) For the paired-end data obtained by sequencing, the sample data is first split according to the Barcode information, and the adapter and Barcode sequence are removed, and then the sequence is spliced ​​to obtain the optimized sequence. After removing the chimera sequence, OUT cluster analysis is performed, and taxonomic analysis is performed on the representative OUT sequence. Based on the OUT cluster analysis results, multiple diversity index analyses and sequence depth detection are performed on OUT; based on the taxonomic information, statistical analysis of colony structure is performed at each classification level.

[0123] 3. Test results

[0124] 3.1. Results of dominant flora in the intestines of maternal mice

[0125] 3.1.1. Changes of Muribaculaceae unclassified in the intestines of female mice

[0126] Changes of Muribaculaceae unclassified in the intestine of female mice Figure 1 As shown, compared with the control group, the relative abundance of Muribaculaceae unclassified in the intestines of pregnant and lactating rats in different doses of N-acetylneuraminic acid intervention groups was significantly higher than that in the control group, and the relative abundance in the high-dose N-acetylneuraminic acid intervention group was higher than that in the low-dose intervention group.

[0127] 3.1.2. Changes of Prevotellaceae UCG-001 in the intestines of female mice

[0128] Changes of Prevotellaceae UCG-001 in the intestines of female mice Figure 2As shown in the data, compared with the control group, the relative abundance of Prevotellaceae UCG-001 in the intestines of pregnant rats in the different doses of N-acetylneuraminic acid intervention groups was higher than that in the control group, and there was no significant difference in the relative abundance between the different doses of intervention groups; the relative abundance of Prevotellaceae UCG-001 in the intestines of lactating rats in the different doses of N-acetylneuraminic acid intervention groups was lower than that in the control group.

[0129] 3.1.3. Changes in Bifidobacterium in the intestines of female mice

[0130] Changes of Bifidobacterium in the intestines of female mice Figure 3 As shown, from before oral gavage to after delivery, the relative abundance of Bifidobacterium in the intestines of pregnant rats in the control group showed a downward trend; compared with the control, the relative abundance of Bifidobacterium in the intestines of pregnant rats in the different doses of N-acetylneuraminic acid intervention groups showed an upward trend, and the high-dose intervention group was higher than the low-dose intervention group; compared with the control, there was no significant difference in the relative abundance of Bifidobacterium in the intestines of lactating rats in the different doses of N-acetylneuraminic acid intervention groups.

[0131] 3.1.4. Changes of Akkermansia in the intestines of female mice

[0132] Changes of Akkermansia in the intestines of female mice Figure 4 As shown, compared with the control group, there was no significant difference in the relative abundance of Akkermansia in the intestines of pregnant rats in the different doses of N-acetylneuraminic acid intervention groups; the relative abundance of Akkermansia in the intestines of lactating rats in the different doses of N-acetylneuraminic acid intervention groups was significantly increased and higher than that of the control group, and the relative abundance of the high-dose intervention group was significantly higher than that of the low-dose intervention group.

[0133] 3.2. Results of dominant bacterial flora in the intestines of offspring mice

[0134] 3.2.1. Changes of Muribaculaceae unclassified in the intestine of offspring mice

[0135] Changes of Muribaculaceae unclassified in the intestine of offspring mice Figure 5As shown, there was no significant difference in the relative abundance of Muribaculaceae unclassified in the intestines of offspring in different groups one week after birth. With the continuation of breast feeding, the relative abundance of Muribaculaceae unclassified in the intestines of offspring in the high-dose N-acetylneuraminic acid intervention group was significantly higher than that in the control group.

[0136] 3.2.2. Changes of Akkermansia in the intestines of offspring mice

[0137] Changes of Akkermansia in the intestines of offspring mice Figure 6 As shown in the data, there was no significant difference in the relative abundance of Akkermansia in the intestines of offspring mice in different groups one week after birth. With the continuation of breastfeeding, the relative abundance of Akkermansia in the intestines of offspring mice in different doses of N-acetylneuraminic acid intervention groups was significantly higher than that in the control group, and the relative abundance of Akkermansia in the high-dose N-acetylneuraminic acid intervention group was higher than that in the low-dose intervention group.

[0138] 3.2.3. Changes of Romboutisa in the intestines of offspring mice

[0139] Changes of Romboutisa in the intestines of offspring mice Figure 7 As shown, one week after the offspring were born, the relative abundance of Romboutisa in the intestines of the offspring in the different doses of N-acetylneuraminic acid intervention groups was higher than that in the control group; with the continuation of breast feeding, the relative abundance of Romboutisa in the intestines of the offspring in the high dose of N-acetylneuraminic acid intervention group was significantly higher than that in the control group.

[0140] 3.2.4. Changes of Escherichia-Shigella in the intestines of offspring mice

[0141] Changes of Escherichia-Shigella in the intestines of offspring mice Figure 8As shown in the data, compared with the control group, the relative abundance of Eschrichia-Shigella in the intestine of the offspring in the high-dose N-acetylneuraminic acid intervention group was lower than that in the control group one week after birth; with the continuation of breast feeding, the relative abundance of Escherichia-Shigella in the intestine of the offspring 3 weeks after birth was significantly reduced, and the relative abundance of Escherichia-Shigella in the intestine of the offspring in the different doses of N-acetylneuraminic acid intervention groups was significantly lower than that in the control group, and the relative abundance in the high-dose intervention group was significantly lower than that in the low-dose intervention group.

Claims

1. Use of N-acetylneuraminic acid substances in the preparation of foods that are ingested by mothers and help regulate the intestinal flora of mothers and their offspring; wherein: The N-acetylneuraminic acid substance is N-acetylneuraminic acid anhydrate, N-acetylneuraminic acid hydrate or N-acetylneuraminic acid salt, the mother is in pregnancy and / or lactation period, and the offspring is in fetal period, infant period, and / or childhood period.

2. The use according to claim 1, characterized in that The method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Muribaculaceae unclassified in the intestinal tract of the pregnant mother, increasing the relative abundance of Prevotellaceae UCG-001 in the intestinal tract of the pregnant mother, and increasing the relative abundance of Bifidobacterium in the intestinal tract of the pregnant mother.

3. The use according to claim 1 or 2, characterized in that The method of helping to regulate the intestinal flora of the mother and her offspring includes: increasing the relative abundance of Muribaculaceae unclassified in the intestinal tract of the lactating mother and increasing the relative abundance of Akkermansia in the intestinal tract of the lactating mother.

4. The use according to any one of claims 1 to 3, characterized in that The method helps to regulate the intestinal flora of the mother and her offspring, including: increasing the relative abundance of Muribaculaceae unclassified in the offspring's intestine, increasing the relative abundance of Akkermansia in the offspring's intestine, increasing the relative abundance of Romboutisa in the offspring's intestine, and reducing at least one of the relative abundance of Escherichia-Shigella in the offspring's intestine.

5. The use according to any one of claims 1 to 4, characterized in that: The food helps to regulate the intestinal flora of the mother and her offspring, thereby affecting the gut-brain axis of the mother and her offspring and thus helping to maintain the health of the nervous system of the mother and her offspring.

6. The use according to any one of claims 1 to 5, characterized in that: The food is infant food, children's food, adolescent food or adult food.

7. The use according to any one of claims 1 to 6, characterized in that The food is a beverage, candy, dairy product, bakery product or dietary supplement.

8. The use according to any one of claims 1 to 7, characterized in that At room temperature, the food is in liquid form, solid-liquid mixed form, solid block form or solid powder form.

9. The use according to any one of claims 1 to 8, characterized in that The food contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional added ingredients and any auxiliary materials acceptable in food.

10. The use according to any one of claims 1 to 9, characterized in that The mass content of the N-acetylneuraminic acid substances in the food is 0.03% to 2.0%.

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